School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, People's Republic of China.
Anal Sci. 2023 Sep;39(9):1541-1550. doi: 10.1007/s44211-023-00370-0. Epub 2023 May 25.
Alkaline phosphatase (ALP) is an essential enzyme involved in cell phosphorus metabolism. Developing sensitive and accurate ALP quantitative assays is significant. In this study, a turn-on chemiluminescence (CL) analysis platform for the detection of ALP activity in human serum was established based on two-dimensional (2D) Fe-centered metal-organic frameworks with 1,3,5-benzene tricarboxylic acid as ligands (denoted as 2D Fe-BTC). The 2D Fe-BTC as the signaling probe reacts with ascorbic acid forming reduced Fe-BTC which catalyzes the luminol CL reaction producing a strong CL signal. The 2D Fe-BTC-based luminol CL reaction exhibited good CL responses when the concentration of ascorbic acid was in the range of 5-500 nM. By employing magnesium ascorbyl phosphate (MAP) as the substrate which can be hydrolyzed by ALP to generate ascorbic acid, a turn-on CL assay for the detection of ALP was established. Under optimal conditions, as low as 0.00046 U L of ALP could be sensitively detected with a linear range of 0.001-0.1 U L. ALP in human serum can be detected after a simple dilution process without any other pretreatment.
碱性磷酸酶(ALP)是参与细胞磷代谢的重要酶。开发敏感和准确的 ALP 定量分析方法具有重要意义。本研究基于以 1,3,5-苯三甲酸为配体的二维(2D)Fe 中心金属有机骨架(表示为 2D Fe-BTC),建立了一种用于检测人血清中 ALP 活性的基于化学发光(CL)的分析平台。2D Fe-BTC 作为信号探针与抗坏血酸反应形成还原的 Fe-BTC,该还原的 Fe-BTC 可以催化鲁米诺 CL 反应,产生强的 CL 信号。基于 2D Fe-BTC 的鲁米诺 CL 反应在抗坏血酸浓度范围为 5-500 nM 时表现出良好的 CL 响应。通过使用镁抗坏血酸磷酸酯(MAP)作为底物,ALP 可以将其水解生成抗坏血酸,建立了一种用于检测 ALP 的基于化学发光的分析方法。在最佳条件下,ALP 的检测下限低至 0.00046 U/L,线性范围为 0.001-0.1 U/L。经过简单的稀释过程,无需任何其他预处理,即可检测人血清中的 ALP。